ChromAtography
UNOsphere™ S Cation
Exchange Support
n
n
n
n
U
ltrahigh binding
capacities at fast linear velocities
H
ydrophilic polymeric
beads engineered
for high mechanical
stability and low
backpressures
R
obust polymer
designed to withstand
repetitive clean-in-
place cycles
B
iopharmaceutical
manufacturing
quantities available
F
ully supported for
regulatory submission
Achieve High Productivity Using UNOsphere S
Cation Exchange Support
Be Productive
In the bioprocess industry, the isolation of
biomolecules from crude feedstock is one of
the most demanding chromatographic steps
in the downstream process.
Biopharmaceutical manufacturers are under
increasing economic pressure to reduce drug
production costs.
These factors require the media used in
the capture step to have very high binding
capacities at fast linear velocities while
maintaining low column backpressure.
UNOsphere is a patented* new-generation
polymeric support, based on a single-step
polymerization process that delivers high
productivity in the capture step.
UNOsphere Polymer Technology
The genesis of UNOsphere support is based
on the single-step polymerization process
used to prepare UNO® continuous-bed
columns. Incorporation of the sulfonic acid
ligand into the matrix during polymerization
leads to consistent batch-to-batch
reproducibility. UNOsphere beads are
macroporous (>2,000 Å), leading to fast
binding kinetics and high binding capacities
(Table 1). Careful selection of monomers and
crosslinkers provides unrivaled base stability
and bead rigidity.
* US patent 6,423,666 B1.
Properties of UNOsphere S Support
Most production chromatography systems have
maximum pressure limits of 3 bar. The median
particle size of UNOsphere S support is 80 µm,
which generates a backpressure less than
1.5 bar at 1,200 cm/hr (Figure 1). The highly
macroporous nature of UNOsphere S support
provides high binding capacities that range from
30–60 mg IgG/ml support in the linear velocity
range of 150–1,200 cm/hr (Figure 1).
Harsh conditions, such as clean-in-place and
corrosive buffer systems, may affect the
long-term stability of chromatographic media.
The robustness of UNOsphere S support
allows it to survive these conditions with
minimal loss of performance (Table 1).
70
1.40
60
1.20
50
40
30
1.00
0.80
0.60
20
10
0
0
200
400
600
800
0.40
0.20
0.0
1,000 1,200 1,400
Linear velocity, cm/hr
Fig. 1. Binding and backpressure properties of
UNOsphere S support. Column size, 1.1 x 20 cm; sample,
2 mg/ml human IgG; buffer, 50 mM sodium acetate, pH 5.0.
(—), backpressure; (—), 10% breakthrough capacity.
Pressure, bar
n
E
fficient capture of
biopharmaceutical
molecules from crude feedstreams
Protein binding capacity, mg
n
Table 1. Properties of UNOsphere S support.
Functional group
–SO3–
Total ionic capacity
280 ± 30 µeq/ml
Dynamic binding capacity*
150 cm/hr
60 mg IgG/ml
1,200 cm/hr
30 mg IgG/ml
Median particle size
80 µm
Recommended linear flow rate range**
50–1,200 cm/hr
Chemical stability
1.0 M NaOH (20°C)
1.0 M HCl (20°C)
Up to 2,000 hr
Up to 200 hr
pH stability range
pH 1–14
Regeneration conditions
1–2 M NaCl
Storage conditions
20% Ethanol
0.1 N NaOH for 30 days
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31
1.0
7
6
5
4
pH
Strong cation
A 280
Type of ion exchanger
0.5
3
2
1
* 10% breakthrough capacity determined with 4.5 mg/ml human IgG in a
1.1 x 10 cm column.
**UNOsphere S support packed into a 20 cm bed height and run at 1,200 cm/hr generates less than 2 bar backpressure.
0.0
0
10
Technical Assistance
Regulatory Support Files are available upon request.
Bio-Rad Laboratories is an ISO 9001 registered
corporation. For additional information and technical
assistance, contact your local Bio-Rad office. In the USA
and Canada, call 1-800-4-BIORAD.
Visit us on the Web at www.bio-rad.com/unosphere/
for more information on Bio-Rad’s complete line of
process chromatography media.
For more information about UNOsphere S support,
request bulletins 2678, 2774, 2780, and 2849. For more
information about the chromatographic performance of
UNOsphere support, refer to the Bibliography. (Note that
the authors refer to UNOsphere as BRX).
30
0
Run time, min
Fig. 2. Purification of murine IgG1 on UNOsphere S column.
Column size, 0.5 x 10 cm (2 ml); sample, 15 ml (6.6 mg) of murine IgG1conditioned medium. The sample was loaded onto the column in 20 mM
citrate-phosphate buffer, pH 4.0, washed, and eluted in a linear gradient
of 0–100% 20 mM citrate-phosphate, pH 8.0 in 10 column volumes at a
flow rate of 2.0 ml/min (600 cm/hr). Each fraction was 2.0 ml. (—), A280;
(—), buffer pH.
1
2
3
4
5
6
7
8
9
150
100
75
50
MW, kD
Capture Performance
UNOsphere S support is designed for high-efficiency
capture of monoclonal antibodies from crude
feedstreams. Murine IgG1 (6.6 mg) was captured and
eluted from a 2 ml UNOsphere S column (Figures 2
and 3); assays of the load and eluate demonstrated
a recovery of 97%. No antibody was detected in the
flowthrough (Figure 3, lane 2). The 10% breakthrough
capacity for this murine IgG1 antibody is 12.8 mg/ml
(at 600 cm/hr) from conditioned medium.
20
37
25
15
10
Fig. 3. SDS-PAGE gel of UNOsphere-purified murine IgG1. Fractions
from the chromatography run shown in Figure 2 were separated on a
4–20% precast gel. On left, reference standards; lane 1, conditioned
medium; lane 2, flowthrough; lanes 3–8, fractions 25, 26, 27, 28, 29, and
30; lane 9, protein A-purified murine IgG1 from culture medium.
Bibliography
Hunter AK and Carta G, Protein adsorption on novel acrylamido-based
polymeric ion exchangers. I. Morphology and equilibrium adsorption,
J Chromatogr A 897, 65–80 (2000)
Hunter AK and Carta G, Protein adsorption on novel acrylamido-based
polymeric ion exchangers. II. Adsorption rates and column behavior,
J Chromatogr A 897, 81–97 (2000)
Ordering Information
Catalog #
Description
156-0111
156-0113
156-0115
156-0117
UNOsphere S Support, 25 ml
UNOsphere S Support, 100 ml
UNOsphere S Support, 500 ml
UNOsphere S Support, 10 L
Bio-Rad
Laboratories, Inc.
Web site www.bio-rad.com USA 800 4BIORAD Australia 61 02 9914 2800 Austria 01 877 89 01 Belgium 09 385 55 11 Brazil 55 21 3237 9400
Canada 905 364 3435 China 86 21 6426 0808 Czech Republic 420 241 430 532 Denmark 44 52 10 00 Finland 09 804 22 00 France 01 47 95 69 65
Germany 089 318 84 0 Greece 30 210 777 4396 Hong Kong 852 2789 3300 Hungary 36 1 455 8800 India 91 124 4029300 Israel 03 963 6050
Italy 39 02 216091 Japan 03 6361 7000 Korea 82 2 3473 4460 Mexico 52 555 488 7670 The Netherlands 0318 540666 New Zealand 0508 805 500
Norway 23 38 41 30 Poland 48 22 331 99 99 Portugal 351 21 472 7700 Russia 7 495 721 14 04 Singapore 65 6415 3188 South Africa 27 861 246 723
Spain 34 91 590 5200 Sweden 08 555 12700 Switzerland 061 717 95 55 Taiwan 886 2 2578 7189 United Kingdom 020 8328 2000
Life Science
Group
Bulletin 2669 Rev B
US/EG
08-0346
0408
Sig 0308